• 제목/요약/키워드: Protein phosphatase

검색결과 901건 처리시간 0.047초

실험적 간 발암모델에서 감마선 조사 돼지고기 섭취가 전암성병변의 생성, 약물대사 효소계 및 소포체 막 안정성에 미치는 영향 (Effects of $\Upsilon$-Irradiated Pork Feeding on Preneoplastic Hepatic Lesion, Cytochrome P450 System and Microsome Glucose 6-Phosphatase Activity in Rat Hepatocarcinogenesis)

  • 김미정;김정희
    • Journal of Nutrition and Health
    • /
    • 제35권6호
    • /
    • pp.643-649
    • /
    • 2002
  • This study was done to investigate effects of ${\gamma}$-irradiated pork feeding on the formation of glutathione S-transferase placental form positive (GST-P$^{+}$) foci, lipid peroxidation, cytochrome P450 system and microsomal glucose 6-phosphatase activity in diethylnitrosamine (DEN)-initiated rat hepatocarcinogenesis. Weaning Sprague-Dawley male rats were fed the diet containing ${\gamma}$-irradiated ground pork at the dose of 0, 3, 10, 30 kGy as a 20% of protein source for 8 weeks. One week after feeding, rats were intraperitoneally injected twice with a dose of DEN (50 mg/kg BW). As a promote.,0.05%phenobarbital was fed in drinking water from one week after DEN treatment until the end of experiment. At the end of 8th week, rats were sacrificed and hepatic GST-P$^{+}$ foci, microsomal malondialdehyde (MDA) and conjugated diene contents were determined. In addition, cytochrome P450 content and the activities of NADPH cytochrome P450 reductase and glucose 6-phosphatase were also measured. There was no significant effect by gamma irradiation on microsomal MDA content, conjugated diene, cytochrome P450 content and activities of NADPH cytochrome P450 reductase and glucose 6-phosphatase. However with DEN treatment, microsomal MDA content showed a increasing tendency. Cytochrome P450 content was also significantly increased while microsomal glucose 6-phophatase activity was significantly decreased with DEN treatment. However the activity of NADPH cytochrome P450 reductase was not affected. An interesting finding in this study was that the number and area of hepatic GST-P$^{+}$ foci of rats fed gamma irradiated pork were tended to be decreased by high dose of irradiation, but were not significantly different. These results might imply that the consumption of low dose of gamma irradiated pork does not affect the formation of hepatic GST-P$^{+}$ foci and lipid peroxide and membrane stability.ability.

Expression, Purification and Functional and structural relationship of pyruvate dehydrogenase phosphatase

  • Kim, Young-Mi;Jung, Ki-Hwa
    • 한국응용약물학회:학술대회논문집
    • /
    • 한국응용약물학회 2002년도 창립10주년기념 및 국립독성연구원 의약품동등성평가부서 신설기념 국재학술대회:생물학적 동등성과 의약품 개발 전략을 위한 국제심포지움
    • /
    • pp.236-236
    • /
    • 2002
  • Pyruvate dehydrogenase phosphatase (PDP) is a mitochondrial protein serine/threonine phosphatase that catalyzes the dephosphorylation and concomitant reactivation of the pyruvate dehydrogenase componant of the pyruvate dehydrogenase complex (PDC). PDP consists of a Mg$\^$+2/ -dependent and Ca$\^$+2)-stimulated catalytic subunit (PDPc) of Mr 52,600 and a FAD-containing regulatory subunit (PDPr) of Mr 95.600. Catalytic subunit of pyruvate dehydrogenase phosphatase (PDPc) has been suggested to have three major functional domains such as dihydrolipoamide acetyltransferase(E$_2$)-binding domain, regulatory subunit of PDP(PDPr)-binding domain, and calcium-binding domain. In order to identify functional domains, recombinant catalytic subunit of pyruvate dehydrogenase phosphatase (rPDPc) was expressed in E. coli JM101 and purified to near homogeneity using the unique property of PDPc: PDPm binds to the inner lipoyl domain (L$_2$) of E$_2$ of pyruvate dehydrogenase complex (PDC) in the presence of Ca$\^$+2/, not under EGTA. PDPc was limited-proteolysed by trypsin, chymotrypsin, Arg-C, and elastase at pH7.0 and 30$^{\circ}C$ and N-terminal analysis of the fragment was done. Chymotrypsin, trypsin, and elastase made two major framents: N-terminal large fragment, approx. 50kD and C-terminal small fragment, approx. 0 kDa. Arg-C made three major fragments: N-terminal fragment, approx. 35 kD, and central fragment, approx. 15 kD, and C-terminal fragment, approx. 10 kD. This study strongly suggest that PDPc consists of three major functional domains. However, further study should be necessary to identify the functional role.

  • PDF

단백질과 마그네슘 공급수준이 흰쥐의 혈청중 지질함량 및 효소활성에 미치는 영향 (Effect of Dietary Protein and Magnesium Levels on Serum Lipid Contents and Enzyme Activities in Rats)

  • 정복미
    • Journal of Nutrition and Health
    • /
    • 제26권8호
    • /
    • pp.925-932
    • /
    • 1993
  • To study the effects of dietary protein and magnesium level on serum lipid contents and enzyme activities, male Sprague-Dawley rats were divided into six groups. Six groups of animals were fed six experimental diets different with protein(8% and 20%) and magnesium levels (Mg-free, 400mg/kg diet and 800mg/kg diet). The serum lipid contents and enzyme activities of rats fed different dietary protein and magnesium levels were examined. Serum triglyceride contents was significantly decreased as the dietary magnesium level was increased. Serum phospholipid contents was significantly higher in magnesium deficient group than those in the other groups. Serum alkaline phosphatase activity was significantly decreased in magnesium dificient group compared with other groups, but serum glutamic oxaloacetic transaminase and glutamic pyruvic transaminase activities was significantly increased in magnesium dificient group than other groups. In conclusion, serum triglyceride contents in low protein level was remarkably decreased as the dietary magnesium level was increased.

  • PDF

식이 단백질에 따른 칼슘수준이 성장기 흰쥐의 체내 칼슘 및 골격 대사에 미치는 영향 (The Effects of Dietary Protein and Calcium Levels on Calcium and Bone)

  • 장영은
    • Journal of Nutrition and Health
    • /
    • 제30권3호
    • /
    • pp.266-276
    • /
    • 1997
  • To investigate the effects of dietary protein and calcium levels on calcium and bone metabolism Sprague-Dawley male growing rats weighting approximately 91.4g were divided into four groups and fed one of the following four experimental diets-15% protein 0.2% calcium ; 15% protein 0.5% calcium ; 30% protein 0.2% calcium ; 30% protein 0.5% calcium-for five weeks. Calcium intake and excretion, apparent calcium absorption were measured and bone densities and mineral contents of femur and scapula were analyzed. Calcium excretion through feces and urine was significantly greater in animals receiving diets of higher calcium. Fecal calcium but not urinary calcium excretion was greater when the protein level was increased from 15% to 30%. Apparent calcium absorption rate was significantly higher with lower calcium intakes. Serum alkaline phosphatase activity was significantly higher in 0.2% calcium group than in 0.5% calcium group, while urinary hydroxyproline excretion was essentially same among all experimental groups. Weights and mineral contents or protein. Bone weights were greater, but calcium and ash contents of femur and scapula were lower in animals on the diet containing low calcium and high protein, which suggests that bone metabolism may be affected by the interaction between calcium and protein intake. These results indicate that during growth high protein intake might be beneficial to bone health if the diet is sufficient in calcium, however, if the diet fails to provide an optimum amount of calcium, such practice might be detrimental.

  • PDF

지방조직 유래 줄기세포의 조골세포로의 분화에 대한 실험적 연구 (A STUDY ON THE OSTEOGENIC DIFFERENTIATION OF ADIPOSE-DERIVED ADULT STEM CELL)

  • 이의석;장현석;권종진;임재석
    • Maxillofacial Plastic and Reconstructive Surgery
    • /
    • 제30권2호
    • /
    • pp.133-141
    • /
    • 2008
  • Stem cells have self-renewal capacity, long-term viability, and multiline age potential. Adult bone marrow contains mesenchymal stem cells. Bone marrow-derived mesenchymal stem cells (BMSCs) are progenitors of skeletal tissue components and can differentiate into adipocytes, chondrocytes, osteoblasts, and myoblasts in vitro and undergo differentiation in vivo. However, the clinical use of BMSCs has presented problems, including pain, morbidity, and low cell number upon harvest. Recent studies have identified a putative stem cell population within the adipose tissue. Human adipose tissue contains pluripotent stem cells simillar to bone marrow-derived stem cells that can differentiate toward the osteogenic, adipogenic, myogenic, and chondrogenic lineages. Human adipose tissue-derived stem cells (ATSCs) could be proposed as an alternative source of adult bone marrow stem cells, and could be obtained in large quantities, under local anesthesia, with minimal discomfort. Human adipose tissue obtained by liposuction was processed to obtain ATSCs. In this study, we compared the osteogenic differentiation of ATSCs in a specific osteogenic induction medium with that in a non-osteogenic medium. ATSCs were incubated in an osteogenic medium for 28 days to induce osteogenesis respectively. Osteogenic differentiation was assessed by von Kossa and alkaline phosphatase staining. Expression of osteocyte specific bone sialoprotein, osteocalcin, collagen type I and alkaline phosphatase, bone morphogenic protein 2, bone morphogenic protein 6 was confirmed by RT-PCR. ATSCs incubated in the osteogenic medium were stained positively for von Kossa and alkaline phosphatase staining. Expression of osteocyte specific genes was also detected. Since this cell population can be easily identified through fluorescence microscopy, it may be an ideal source of ATSCs for further experiments on stem cell biology and tissue engineering. The present results show that ADSCs have an ability to differentiate into osteoblasts. In the present study, we extend this approach to characterize adipose tissue-derived stem cells.

치주인대세포에 대한 Bone morphogenetic protein-7의 영향 (Effect of BMP-7 on the rat periodontal ligament cell)

  • 김경희;김영준;정현주
    • Journal of Periodontal and Implant Science
    • /
    • 제35권2호
    • /
    • pp.289-298
    • /
    • 2005
  • Bone morphogenetic protein-7(BMP-7), a member of the transforming growth factor superfamily, stimulates osteoblast differentiation and bone formation. There are lots of evidences supporting a direct participation of periodontal ligament(PDL) cells on periodontal tissue regeneration. The purpose of this study was to evaluate the effect of recombinant human(rh) BMP-7 on primary rat PDL cells in vitro, with special focus on the ability of bone formation. The PDL cells were cultured with rhBMP-7 at the concentration of 0, 10, 25, 50, 100 and 200ng/ml for MTT assay. We evaluated the alkaline phosphatase activity at 3 and 5 days of incubation and the ability to produce mineralized nodules of rat PDL cells at 14 days of cell culture in concentration of 0, 10, 25, 50 and 100ng/ml. The cell activity was not reduced in cells treated with BMP-7 at $10{\sim}100ng/ml$, whereas the cell activity was reduced in the concentration of 200ng/ml than the control at day 1 and 3(p<0.01). At 3 and 5 day, alkaline phosphatase activity was significantly increased in cells treated with BMP-7 at 50ng/ml and 100ng/ml(p<0.05). The area of mineralized bone nodule was greater in cells treated with BMP-7 at 50 and 100 ng/ml than the control(p<0.01). These results suggest that rhBMP-7 stimulate rat PDL cells to differentiate toward osteoblast phenotype and secretion of the extracellular matrix of rat PDL cells.